1. Field of the Invention
A present invention discloses a monoclonal antibody against Der p 2, hybridoma cell line producing thereof and test strip, kit and method for dust mite assay.
2. Description of the Related Art
Asthma and allergic rhinitis are the most common airway allergic diseases in Taiwan. Both of them are resulted from the inflammation induced by the interaction between respiratory epithelial cells and allergens. The major allergens include hair of vertebrates, egg, murine urine, poisonous fluid of insect, dust mite, cockroach, shrimp, pollen, peanut and fungal. Therein, the dust mite exhibiting rich and diverse allergens majorly distributes at the food, furniture, interior decoration, bedding, carpet and dust within the indoor environment. Besides, allergens from dust mite are particularly easy to cause allergy of airway and skin. Therefore, dust mite is considered as the major allergen in the interior environment.
Previous reports show that dust mite allergens are divided into 22 groups according to their biological characteristics, molecular weight and binding affinity with immunoglobulin E (IgE). Therein, the major allergens are group 1 and group 2 allergen of Dermatophagoides pteronyssiuns (Der p 1 and Der p 2). The IgE in the serum from 75% of the allergic patients exhibits the binding response to Der p 1 and Der p 2. Previous reports also suggested that reduce the concentration of allergens in the environment, especially at the onset time of asthma and allergic rhinitis, could efficiently prevent the occurrence of the allergic diseases.
Der p 1 belongs to enzymatic allergen which is unstable to be applied as the real time indicator. In contrast, Der p 2, a stable non-enzymatic allergen, which exhibits the great correction with the number of dust mite in the environment, is a capable indicator for detecting the concentration of dust mite allergen at home. Allowing patients to detect the number of dust mite and control the allergen concentration at home environment brings more economic profit than pharmaceutical therapy.
Monoclonal antibody technology has been widely applied for rapid detection kit because of its advantages, including high specificity, fast reaction and high sensitivity. However, there are still many uncertainties during the production of monoclonal antibody. Therefore, finding the monoclonal antibody with high specificity is the critical point in the development of detection kit.
U.S. Pat. No. 6,132,734 disclosed the amino acid sequence of Der p 2, but did not reveal the specific epitope of the Der p 2 or the monoclonal antibody against Der p 2. In addition, Taiwan Pub. No. 200414904 disclosed the method for producing the monoclonal antibody, WAN-108, via injecting the allergen protein isolated by gel electrophoresis and immunoblotting into recipient mice. However, the allergenic type, recognized epitope and antibody activity of WAN-108 monoclonal antibody still remained elusive. Accordingly, these previously disclosed dust mite detection technologies still remain to be improved.